Optimization of Expression Conditions of an Induction Strategy for Improving Liver Targeted Interferon (IFN-CSP) Production in <i>E</i>.<i>coli</i>_Journal of Biomedical Engineering

Authors：

HUANGYanting , LUXuemei , WANGJie , JINXiaobao ,  ZHUJiayong

Institute of Pharmaceutical Bioactive Substances, Guangdong University of Pharmaceutical;Guangdong Key Laboratory of Pharmaceutical Bioactive Substances, Guangzhou 510006, China;

Corresponding author：

ZHUJiayong, Email: zhujy@gdpu.edu.cn

Keywords：

liver targeted interferon; Escherichia coli; opptimal expression condition; orthogonal experimental design

DOI：

10.7507/1001-5515.20140080

Video：

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Expression conditions of induction strategies for the cytoplasmic inclusion bodies (IBs) production of liver targeted interferon IFN-CSP by recombinant Escherichia coli (E.coli) BL21(DE3) were optimized in shake-flask cultures in this study. The factors of the optimized protocol included in the present study were pH, inducer IPTG (isopropyl β-D-thiogalactoside) concentration, culture growth temperature, incubation time and induction point. The effects of those factors were investigated by ‘single variable at a time’ method, aimed to analyze characterization of the recombinant strain. Orthogonal experimental design was further used to optimize the above critical factors for IFN-CSP production. According to the expression optimization result, it was confirmed that the main influence factors were cell density and induction temperature. The IFN-CSP gene expression optimized conditions were:pH value of the culture medium was 6.0, culture temperature 37℃, adding IPTG to final concentration 0.4 mmol/L when the recombinant strain growth density OD₆₀₀ achieved 0.8 and induction time 4 h. At this point, the IBs represented 74.3% of the total cellular protein. Compared with the non-optimized condition, IFN-CSP production obtained in optimized induction strategies were increased by approx. 1.2-fold. The optimized induction strategy yielded 688.8 mg/L of IFN-CSP, providing experimental data to study the biology activity and productive technology of IFN-CSP.

Citation： HUANGYanting, LUXuemei, WANGJie, JINXiaobao, ZHUJiayong. Optimization of Expression Conditions of an Induction Strategy for Improving Liver Targeted Interferon (IFN-CSP) Production in E.coli. Journal of Biomedical Engineering, 2014, 31(2): 432-438. doi: 10.7507/1001-5515.20140080 Copy

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2. KEW M C.Epidemiology of chronic hepatitis B virus infection,hepatocellular carcinoma,and hepatitis B virus-induced hepatocellular carcinoma[J].Pathol Biol (Paris),2010,58(4):273-277.
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10. LI M,HUANG D.Purification and characterization of prokaryotically expressed human interferon-lambda2[J].Biotechnol Lett,2007,29(7):1025-1029.
11. LI Hui,LIU Qingyong,CUI Kuiqing,et al.Expression of biologically active human interferon alpha 2b in the milk of transgenic mice[J].Transgenic Res,2013,22(1):169-178.
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17. AZAMAN S N,RAMAKRISHNAN N R,TAN J S,et al.Optimization of an induction strategy for improving interferon-alpha2b production in the periplasm of Escherichia coli using response surface methodology[J].Biotechnol Appl Biochem,2010,56(4):141-150.

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Optimization of Expression Conditions of an Induction Strategy for Improving Liver Targeted Interferon (IFN-CSP) Production in E.coli

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